Volume 11, Issue 44 (9-2021)                   NCMBJ 2021, 11(44): 95-106 | Back to browse issues page

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Abbasifarid E, Bolhassani A, Irani S, Sotoodehnejadnematalahi F. Design of Eukaryotic Vector Harboring L1-E7 Polyepitope Fusion Construct of High Risk Human Papillomaviruses. NCMBJ. 2021; 11 (44) :95-106
URL: http://ncmbjpiau.ir/article-1-1414-en.html
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Abstract:   (591 Views)
Abstract:
Aim and Background: Human Papillomaviruses (HPV) infection was known to be the leading cause of cervical cancer. Cervical cancer is the fourth most common cancer among women in the world. Designing a DNA vaccine with therapeutic properties as well as prevention can have a significant impact on reducing morbidity and mortality.
Materials and Methods: Gene sequences including the immunogenic and conserved epitopes of L1 and E7 proteins of high risk papillomaviruses were designed. After synthesis of the L1-E7 fusion construct in pUC57 cloning vector, its subcloning was performed in pcDNA3.1 (-) eukaryotic vector. The concentration and purity of the recombinant pcDNA-L1E7 plasmid was determined by NanoDrop spectrophotometry.  
Results: The gene sequence after cutting from pUC57-L1E7 by BamHI/HindIII restriction enzymes, was subcloned in the same cloning site of pcDNA3.1 (-) vector. The presence of gene was confirmed by digestion with BamHI/HindIII enzymes, as a 639 bp fragment on 1% agarose gel. The recombinant pcDNA-L1E7 plasmid was purified by endotoxin-free extraction kit for isolation of bacterial lipopolysaccharide. The concentration of the purified DNA was obtained about 1474 ng/ µl.
Conclusion: Cloning of the L1-E7 fusion construct in the eukaryotic vector was successful. In the next steps, the recombinant vector will be used for gene vaccine studies in vivo
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Type of Study: Research Article | Subject: Cellular and molecular
Received: 2021/04/14 | Accepted: 2021/07/23 | Published: 2021/09/1

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